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Glucosyl stevia composition

Active Publication Date: 2014-08-14
PURECIRCLE SDN BHD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is about improving Stevia sweeteners by producing a high purity food ingredient from the extract of the Stevia rebaudiana Bertoni plant. The ingredient contains glucosylated derivatives of stevioside, Rebaudioside A, Rebaudioside B, Rebaudioside C, Rebaudioside D, Rebaudioside E, Rebaudioside F, dulcoside A, steviolbioside, rubusoside, and other steviol glycosides found in the plant. The process involves using an enzymatic transglucosylating process with CGTases produced by cultures of Bacillus stearothermophilus. The process can also include steps of decolorizing, desalting, and removing maltooligosaccharides and unmodified (unreacted) steviol glycosides. The ingredient can be used as a sweetener and flavor modifier in various foods and beverages.

Problems solved by technology

However many artificial sweeteners such as dulcin, sodium cyclamate and saccharin were banned or restricted in some countries due to concerns on their safety.
However, even in a highly purified state, steviol glycosides still possess undesirable taste attributes such as bitterness, sweet aftertaste, licorice flavor, etc.
One of the main obstacles for the successful commercialization of stevia sweeteners are these undesirable taste attributes.
However in such process, the resulting product contains high level of initial unreacted (unmodified) glycosides (generally >20%) which makes it not compliant with regulatory requirements of less than 15% unreacted glycosides (α-Glucosyltransferase Treated Stevia, Japan's Specifications and Standards for Food Additives, VIII edition, 2009, p.
However chromatographic separation techniques generally involve high cost and are not suitable for large scale production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of CGTase

[0072]A strain of Bacillus stearothermophilus St-100 was inoculated in 2,000 liters of sterilized culture medium containing 1.0% starch, 0.25% corn extract, 0.5% (NH4)2SO4, and 0.2% CaCO3 (pH 7.0-7.5) at 56° C. for 24 hrs with continuous aeration (2,000 L / min) and agitation (150 rpm). The obtained culture broth was filtered using Kerasep 0.1 μm ceramic membrane (Novasep, France) to separate the cells. The cell-free permeate was further concentrated 2-fold on Persep 10 kDa ultrafilters (Orelis, France). The activity of the enzyme was determined according to Hale, Rawlins (1951). A crude enzyme preparation with activity of about 2 unit / mL was obtained.

example 2

Preparation of β-Amylase

[0073]A strain of Bacillus polymyxa St-3504 was inoculated in 2,000 liters of sterilized culture medium containing 1.0% starch, 0.5% peptone, 0.5% corn extract, 0.5% NaCl, 0.02% MnSO4 and 0.1% CaCO3 (pH 7.0-7.5) at 32° C. for 24 hrs with continuous aeration (2,000 L / min) and agitation (150 rpm). The obtained culture broth was filtered using Kerasep 0.1 μm ceramic membrane (Novasep, France) to separate the cells. 10% of glucose was added to the cell-free permeate which was further concentrated on Persep 10 kDa ultrafilters (Orelis, France) and dried using Alpha 1-4 LSC freeze drier unit (Christ, Germany) to obtain a powder with 20,000 AUN / g activity. β-Amylase activity unit (1 AUN) was defined as the activity which liberates 100 mg of reducing sugar (expressed by dextrose equivalent) per minute under the following conditions: 1 mL of enzyme solution is mixed with 5 mL of 1.2% starch solution (pH 5.5, M / 20 Acetate Buffer) and kept for 20 min at 40° C.

example 3

Preparation of Short-Chain Glucosyl Stevia Composition

[0074]100 g of tapioca starch was suspended in 300 mL of water (pH 6.5). 2 KNU of α-amylase (Termamyl Classic, Novozymes, Denmark) and 30 units of CGTase obtained according to EXAMPLE 1 were added, and the liquefaction of starch was carried out at 80° C. for about one hour to dextrose equivalent about 15. The pH of reaction mixture was adjusted to pH 2.8 by hydrochloric acid and the mixture was boiled at 100° C. during 5 minutes to inactivate the enzymes. After cooling to 65° C., the pH was adjusted to pH 6.0 with sodium hydroxide solution. 100 g stevia extract produced by PureCircle (JiangXi) Co., Ltd. (China), containing stevioside 29.2%, Rebaudioside A 54.3%, Rebaudioside C 9.0%, Rebaudioside F (1.7%) and other glycosides amounting to total steviol glycosides content of about 96.4% was added to liquefied starch and stirred until a homogeneous solution was obtained. 200 units of CGTase was added to the solution and the mixture ...

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Abstract

Glucosyl stevia compositions are prepared from steviol glycosides of Stevia rebaudiana Bertoni. The glucosylation was performed by cyclodextrin glucanotransferase using the starch as source of glucose residues. The short-chain glucosyl stevia compositions were purified to >95% content of total steviol glycosides. The compositions can be used as sweetness enhancers, flavor enhancers and sweeteners in foods, beverages, cosmetics and pharmaceuticals.

Description

RELATED APPLICATIONS[0001]This application is a continuation-in-part application of and claims the benefit of priority from:[0002]co-pending U.S. patent application Ser. No. 14 / 073,423, filed on Nov. 6, 2013, which is a continuation application of U.S. patent application Ser. No. 13 / 567,707, filed on Aug. 6, 2012 and issued as U.S. Pat. No. 8,647,844 on Feb. 11, 2014, which is a divisional application of U.S. patent application Ser. No. 13 / 029,263, filed on Feb. 17, 2011 and issued as U.S. Pat. No. 8,257,948 on Sep. 4, 2012; and[0003]co-pending U.S. patent application Ser. No. 13 / 589,754, filed on Aug. 20, 2012, which is a continuation of U.S. patent application Ser. No. 13 / 074,179, filed on Mar. 29, 2011 and issued as U.S. Pat. No. 8,318,459 on Nov. 27, 2012, which is a continuation-in-part application of U.S. patent application Ser. No. 13 / 029,263, filed on Feb. 17, 2011 and issued as U.S. Pat. No. 8,257,948 on Sep. 4, 2012.[0004]The contents of each of these applications are inco...

Claims

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Application Information

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IPC IPC(8): A23L1/221A23L27/10
CPCA23L1/221A23C9/1307A23L2/60A21D2/18A21D2/36A61K36/28C12P19/18C12P19/56C12P33/00A61K31/704A23L2/02C12P19/22C12Y204/01019A23L27/36A23L2/54Y02P20/52A23L27/33A23L33/105A23V2002/00
Inventor MARKOSYAN, AVETIK
Owner PURECIRCLE SDN BHD
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